Aldosterone increases VEGF-A production in human neutrophils through PI3K, ERK1/2 and p38 pathways.

Abstract Aldosterone is now recognised as an important actor in inflammation processes. Neoangiogenesis plays a crucial role in this complex process and immune cells, such as neutrophils, appear to be able to secrete different forms of (pro)angiogenic molecules, especially VEGF-A. The present work was undertaken to investigate whether aldosterone was able to regulate VEGF-A production in human neutrophils. The HL-60 (progranulocytic) cell line and human polymorphonuclear leukocytes were incubated for different time periods with aldosterone. Total cellular RNA extraction, submitted to reverse transcription and real time semi-quantitative PCR, was used to study VEGF-A mRNA expression. Cell supernatants were collected and ELISA tests were performed to analyse VEGF-A protein production. Aldosterone increased VEGF-A mRNA and protein expression in a dose- and time-dependent manner in both cell types. Inhibitors of PI3 kinases, ERK1/2, and to a lesser extent of p38 MAPK, decreased this aldosterone-induced immune cell activation. Western-blot performed with HL-60 cells confirmed that ERK1/2 and p38 MAPK pathways were stimulated by aldosterone. Mineralocorticoid receptors are implicated in this VEGF-A up-regulation because HL-60 cells pre-treated with spironolactone, an aldosterone receptor antagonist, diminished the effects of aldosterone. Aldosterone was also able to increase VEGF-A production of phagocytic cells such as neutrophils. These results suggest that this hormone could play an active role in the neovascularisation process by favouring entry of plasma proteins and fluids into the vascular wall, cell proliferation and tissue rebuilding.